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Objective@#By analysis the differences of technical specifications among National Health Commission of the People’s Republic of China, Centers for Disease Control and Prevention (CDC) and European Network of Infectious Disease (EUNID)domestic and foreign, to provide a reference for the construction of high-level isolation units (HLIUs) in China.
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Objective@#To evaluate, the method of different packing layer and sealing mode for the packaging of the infectious wastes in order to achieve better autoclave sterilization effect.@*Methods@#In the autoclave cycles P4 and P9, different packing layer (single, double and triple) and sealing mode (slightly folded, autoclave tape, nylon tape closure) were applied to the internal infectious waste sterilization, biological indicators were used to detect the effect of autoclave sterilization of the upper, middle and lower layers, respectively.@*Results@#1) In the autoclave cycle P4, the triple autoclave bags packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper layer, the middle layer and the lower layer were respectively in the upper layer 100%, 0%, 0%, in the middle layer 67%, 20%, 0%, and in the lower layer was 0%. The double autoclave bags packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper layer, the middle layer and the lower layer were respectively the in the upper layer 100%, 85%, 0%, the middle layer 45%, 0%, 0%, and the lower layer 10%, 0%, 0%. the single autoclave bag packaging sealing method wa based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper, middle and lower layers were respectively the upper layer 100%, 100%, 0%, the middle layer 70%, 45%, 0%, and the lower layer 12.5%, 0%, 0%. (2) In the autoclave cycle P9, the triple autoclave bags packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper layer, middle and lower layers were respectively the upper layer all 100%, the middle layer 100%, 100%, 70%, and the lower layer 100%, 100%, 30%. The double autoclave bags packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper, middle and lower layers were respectively the upper layer all 100%, the middle layer all 100% and the lower layer is 100%, 100%, and 60%. The single autoclave bag packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper, middle and lower layers were all 100%.@*Conclusions@#When autoclave is recommended for sterilization of infectious waste, the best effect was achieved with a single unsealed autoclave bag. If it is necessary to seal, the autoclave bag should not be completely sealed, avoiding incomplete penetration of steam into the interior space, otherwise the infectious waste can not be completely sterilized.
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Objective@#To understand the pathogenic composition of hand, foot and mouth disease (HFMD) in Ningxia Hui Autonomous Region (Ningxia) from 2016 to 2017, and analyze the genetic characteristics of the main pathogens enterovirus (EV)-A71 and coxsackievirus (CV)-A16.@*Methods@#Analysis of the result of nucleic acid testing of HFMD in Ningxia from 2016 to 2017 to determine the pathogenic composition of HFMD. The complete VP1 coding region was amplified by RT-PCR and the gene sequence was determined for the enterovirus strains sent to the National HFMD Network Monitoring Laboratory in Ningxia from 2016 to 2017. BLAST analysis confirmed the serotype of the strain, and the phylogenetic tree was constructed respectively by selecting EV-A71 and CV-A16 isolates.@*Results@#The leading pathogens of HFMD in Ningxia of 2016 and 2017 were other EV (397, 43.72%) and EV-A71 (918, 56.18%) respectively, and the dominant pathogens in different months may differ. The pathogenic composition causing HFMD in the past two years has changed from CV-A16 and other EV to EV-A71 and other EV. The isolated EV-A71 strains were C4a evolutionary branch and the isolated CV-A16 strains were B1b evolutionary branch.@*Conclusions@#Compared to 2016, in 2017 EV-A71, CV-A16 and other EV changed dynamically. Dynamic monitoring of EV-A71 in Ningxia is of great significance to guide the strategy of using EV-A71 vaccine, concentrating medical resources to strengthen the treatment and reduce the mortality rate of severe HFMD cases.
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Objective@#To explore the research status of rhinovirus (RV) through analysis of rhinovirus literature using GoPubMed.@*Methods@#"Rhinovirus" was used as the major subject word and the rhinovirus literature was collected at PubMed database (from Jan 1, 1970 to April 16, 2018). The high frequency subject words of rhinovirus related literature and the distribution of countries, cities, and journals were analyzed through a bibliometrical analysis method .@*Results@#A total of 5 367 reports were retrieved from PubMed. The quantity of rhinovirus papers increased overall year by year. The highest number of papers were mainly published in developed countries. The highest number of papers on RV were mainly published in J Virol among all journals related with rhinovirus and Tyrrell D published the highest number of papers in all authors contributed to articles on rhinovirus. The rhinovirus, human, virus, respiratory tract infection were the high frequency subject words in the rhinovirus research.@*Conclusions@#Rhinovirus research is becoming one of research hotspots according to the statistical analysis of the research literature on rhinoviruses by GoPubMed.
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The laboratories of National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, have developed a series of experimental method . These method have unique advantages over the national standard method and industry standard method . How to make these method become public products through legal procedures to serve disease prevention and control more extensively is an obligatory task for national virological laboratories. This article explores the establishment, verification, validation, and examination of virological non-standard method under laboratory certification and accreditation conditions through empirical research on the " RT-RAA method for detecting EV71/CA16 and other viruses" .
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As one part of Overall National Security, Biosafety has became a basic strategy for adhering to and developing the socialism with Chinese characteristics in the new era, which also meets the requirements of national strategies. Since 2004, microbiological laboratory biosafety of China has made significant progress in China, and gave a positive contribution to Ebola fight in Serra Leone in 2014. In the new era, microbiological laboratory biosafety of China should insist on the core problem, cover the total issues and make sure enough measures be in place. Under the principal of national strategy of Implementing Healthy China, the Belt and the Road Initiative, building a community of human destiny, microbiological laboratory biosafety will centre on "3B" problems related to biosafety, biosecurity and biotechnology, and provide necessary guarantee to re-emerging infectious disease control and public health security.
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Objective@#To establish quantitative real-time PCR (qPCR) method based on Taqman probe for detecting Ekpoma virus (EKV).@*Methods@#According to the conserved region of gene in EKV genome from GenBank, primers and probe for qPCR were designed. Validity and sensitivity were evaluated in this study. Both whole blood and serum of a returnee from Angola were tested by the established EKV-1 and EKV-2 qPCR method .@*Results@#Sensitivity of EKV-1 and EKV-2 qPCR method was respectively 41 copies/μl and 70 copies/μl. Coefficient of variance (CV) was respectively 1.27%, 0.20%, 0.82%; 2.12%, 1.74%, and 1.40%. EKV-2 gene was detected in both whole blood and serum of a returnee from Angola.@*Conclusions@#The first EKV-2 gene was confirmed in both whole blood and serum of a returnee from Angola by real-time RT-PCR..
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Objective@#To test the performance of two kinds of protective respirators (3M9332, 3M1860) used in Biosafety Level 3 Laboratory (BSL-3) to ensure whether it can fit well in different populations in order to reduce the risk of infection in the process of operating high-risk pathogenic microorganisms.@*Methods@#Using a 8038PortaCount®Pro+ respirator fit tester to examine the fitness of personnel wear 3M 1860 N95 and 9332 N99 respirators. The influence factors such as gender, age, shape of respirator were evaluated. Statistical analysis was performed by the fit factor(FF) of assigned actions.@*Results@#A total of 62 people conducted the respirator fit test, of which 45 people passed the test of 3M9332 respirators, the pass rate was 72.58%; only 6 people passed the test of 3M1860 respirators, the pass rate was 9.67%. The pass rate of two different types of respirators was analyzed statistically, P=0.000. The P value of assigned actions in deep breathing, head side to side, head up and down, talking, bending over were respectively 0.094, 0.076, 0.542, 0.000, 0.000.@*Conclusions@#The experimental results showed that the type of respirator had a significant effect on respirator fit in this study, but gender and age had no significant effect on factors. As to reduce the risk of leakage of the respirator, redundant actions should try to be avoided, especially loud talking and bending in the course of the experiment.
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Objective To establish an inactivated viral particle-based ELISA for the detection of antibodies against Middle East respiratory syndrome coronavirus (MERS-CoV) in serum samples collected from a MERS-CoV associated case. Methods Serum samples were collected from 10 newborns and 40 healthy adults. A viral particle-based ELISA was established by using the inactivated MERS-CoV virions as antigen. The levels of IgM and IgG antibodies in the serum samples were detected by the established ELISA and the cut-off values for positive detection were determined. Then the inactivated MERS-CoV virion-based ELISA was used to detect the antibodies against MERS-CoV in 5 serum samples collected from the first im-ported MERS case in China. Results The cut-off values of IgM and IgG antibodies in serum samples for ELISA were determined to be A450 readings of 0. 32 and 0. 42, respectively. The titers of IgM and IgG anti-bodies in serum samples collected at early admission to hospital from the first imported MERS case in China were both 1 ︰ 40. Seroconversion occurred 2 weeks after his admission to hospital with the titers of IgM and IgG reaching to 1 ︰ 320. Conclusion The inactivated MERS-CoV virion-based ELISA was established successfully and could be used for the detection of serum antibodies (IgG and IgM) in MERS associated cases.
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Objective To assess the immune level of cervical cancer patients and precancerous lesions patients,and investigate the association of Th 1 /Th2 type cytokines expression and clinical pathologic factors in the plasma of Uyghur cervical cancer patients .Methods we collected peripheral blood specimens from cervical cancer patients,precancerous lesion(CINⅢ)patients and controls.The expressions of Th1 type cytokines IL-2, IFN-γand Th2 type cytokines IL-4,IL-10 in plasma were detected by ELISA .Receiver operating characteris-tic curve( ROC curve) was used to analyze the cytokines value of auxiliary diagnosis in cervical cancer .Results The results showed that compared with control group ,the expressions of Th1 type cytokines IL-2,IFN-γin cer-vical cancer and precancerous lesion group were significantly reduced (P<0.05).The expressions of Th2 type cytokines IL -4, IL -10 in cervical cancer and precancerous lesion group were significantly increased ( P <0.05).in cervical cancer group,IL -10 expression gradually increased with tumor pathological staging (P <0.05).IL-2 expression level in Uyghur cervical cancer patients was significantly lower than in Han patients (P<0.05).Compared cervical cancer group with control group ,AUC of IL-2,IFN-γ,IL-4 and IL-10 were 0.979,0.766,0.736 and 0.903.Conclusion Cellular immune level of cervical cancer patients was low and Th1/Th2 shift has occurred , which suggests that it may be one of the mechanisms in immune escape of tumor cells.Th1 /Th2 type cytokines detection has a certain significance for auxiliary diagnosis of cervical cancer .Be-sides,the decrease of IL-2 expression may play an important role in the occurrence and development of Uyghur patients with cervical cancer .
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The quality control process throughout the Ebola virus nucleic acid detection in Sierra Leone-China Friendship Biological Safety Laboratory (SLE-CHN Biosafety Lab) was described in detail, in order to comprehensively display the scientific, rigorous, accurate and efficient practice in detection of Ebola virus of first batch detection team in SLE-CHN Biosafety Lab. Firstly, the key points of laboratory quality control system was described, including the managements and organizing, quality control documents and information management, instrument, reagents and supplies, assessment, facilities design and space allocation, laboratory maintenance and biosecurity. Secondly, the application of quality control methods in the whole process of the Ebola virus detection, including before the test, during the test and after the test, was analyzed. The excellent and professional laboratory staffs, the implementation of humanized management are the cornerstone of the success; High-level biological safety protection is the premise for effective quality control and completion of Ebola virus detection tasks. And professional logistics is prerequisite for launching the laboratory diagnosis of Ebola virus. The establishment and running of SLE-CHN Biosafety Lab has landmark significance for the friendship between Sierra Leone and China, and the lab becomes the most important base for Ebola virus laboratory testing in Sierra Leone.
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Humans , China , Ebolavirus , Classification , Genetics , Hemorrhagic Fever, Ebola , Diagnosis , Virology , Laboratories , Workforce , Reference Standards , Laboratory Infection , Quality Control , RNA, Viral , Genetics , Sierra LeoneABSTRACT
To investigate the genetic character and origin of the first imported infection case of middle East respiratory syndrome coronavirus (named as MERS-CoV_China GD01), RNA was extracted from swabs of this patient followed by RT-PCR amplification. All coding gene of structural (S, E, M, E) and accessory (ORF3, ORF4a, ORF4b, ORF5, ORF8b) proteins were sequenced and analyzed. Phylogenetic analyses of structural protein coding genes of MERS-CoV_ China GD01 indicates that several substitutes exists in S coding gene and its origin belong group 5 of MERS-CoV, which were recent circulated in Saudi Arabia area, while other three structural genes (N, E, M) were very conserved. Phylogenetic analyses of accessory protein coding genes of MERS-CoV China GD01 indicates that several substitutes exists among ORF3, ORF4a, ORF4b and ORF5, while ORF8b was conserved. In conclusion, genome of MERS-CoV_ China GD01 was general conserved although several genetic variations were found among structural and accessory protein coding genes. This is the first report on sequencing and phylogenetic analyses of the first imported MERS case in China, which may pay the way for prevention and control of imported MERS-CoV infection.
Subject(s)
Humans , China , Conserved Sequence , Coronavirus Infections , Virology , Evolution, Molecular , Genomics , Middle East Respiratory Syndrome Coronavirus , Genetics , Physiology , Phylogeny , Sequence Analysis , Viral Proteins , GeneticsABSTRACT
Objective To establish differential protein expression profile specific for serum proteome of cervical carcinoma in Uighur women by protein 2-dimensional liquid phase chromatogra-phy. Methods We collected and prepared sera of cervical carcinoma from Uighur women, and sepa-rated and processed data using 2-dimensional liquid phase chromatography system specific for protein (ProteomeLabTM PF-2D ). Results A differential expression profile of serum proteome based on characteristics of protein isoelectrie point and hydrophobic features was successfully established. Fifty-six differentially expressed protein spots were found, among which there was an obvious difference in 12 peaks. Conclusion Two-dimensional liquid phase chromatography is an effective and feasible method to establish differential expression profile of serum proteome and to offer a new way for the screening of serum protein markers of Uighur women with cervical carcinoma.
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Objective To establish a high quality genomic DNA preparation method from formalin fixed and paraffin embedded tissues (FFPET) by integrating previous DNA extraction methods. Methods We combined classical genomic DNA extraction methods and commercially available DNA affinity column, replaced the de-waxing by dimethylbenzene with water-bath, designed a fast and improved genomic DNA preparation method. We also extracted genomic DNA from paraffin embedded cervical cancer tissues, and checked the quality of DNA by agarose gel electrophoresis and polymerase chain reaction detection. Results The improved genomic DNA extraction method combined the advantages of the water-bath de-waxing and DNA affinity column, making it possible to get high quality genomic DNA from paraffin embedded cervical cancer tissues, and especially efficient to recover genomic DNA fragments larger than 20 kb. Conclusion The improved DNA extraction method is fast and convenient to recover high quality genomic DNA from paraffin embedded tissues.
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Objective To investigate the influencing factors on re-examination compliance of patients with breast cancer Who were in the stage of recovery.Methods By clinic services and telephones,we investigated the reexamination compliance of 189 cases of breast cancer.Results 66.1% of the cases took periodic re-examinations and on the opposite,33.9% of the patients didn't do so.Statistical meaning could be found from the difference of ages,living sites and survival time limits(P＜0.05).Conclusions The main factors to influence the re-examination compliance of patients with breast cancer are age,living site and survival time limit.And it is possible to improve the re-examination compliance of patients by enhancing health education,raising medicsl levels and service qualities,and creating comfortable environment for patients.